CN105295441A - Stabilizer of chromogenic reagent and applications thereof - Google Patents
Stabilizer of chromogenic reagent and applications thereof Download PDFInfo
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- CN105295441A CN105295441A CN201510811109.8A CN201510811109A CN105295441A CN 105295441 A CN105295441 A CN 105295441A CN 201510811109 A CN201510811109 A CN 201510811109A CN 105295441 A CN105295441 A CN 105295441A
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Abstract
The present invention relates to the field of chemical detection, and in particular relates to a stabilizer of a chromogenic reagent and applications thereof. According to the present invention, an azo dye is used as a stabilizer of a chromogenic reagent, so that a good stabilization effect is exerted and detection is not interfered. Experiments show that, by adding the azo dye, the spontaneous coloration of the chromogenic reagent is improved and the chromogenic reagent maintains better color rendering ability. In a check test without adding the azo dye, a staining agent has color changes to a certain extent, and the light absorption value is also significantly increased. After the stabilizer provided by the present invention is used, color change of the staining agent is not significant, and the light absorption value change is also not significant. Moreover, the stabilizer placed at high temperature for a long period still has a good detection effect.
Description
Technical field
The present invention relates to field of chemical detection, particularly relate to a kind of stablizer and application thereof of developer.
Background technology
At present, to mensuration (qualitative, the quantitative) method of target component, mostly apply the redox reaction that enzyme participates in.The method can be carried out according to following pattern usually: such as, generates oxidizing substance by target component to be measured, utilizing oxydase that this oxidizing substance is reacted with by being oxidized the developer generating color-developing compounds, measuring the absorbancy of the colour developing produced.In the method, the size of absorbancy is suitable with the amount of the color-developing compounds generated, and the amount of the aforementioned color-developing compounds generated is suitable with the amount of the oxidizing substance generated, and the amount of aforementioned oxidation material is suitable with the amount of target component.That is, by detecting the colour developing (color-developing compounds generated) produced, indirectly target component can be measured by such redox reaction.
As mentioned above, developer is very extensive in clinical biochemical detection application.But there is the developer of redox active, often very unstable.Usually occur due to add enzyme and the artificial redox reaction that causes start before, namely produce nature when preserving and to develop the color such problem.Its reason be presumably due to: such as when aforementioned wet system, in aforementioned solvents, aforementioned developer becomes unstable, and when responsibility, owing to absorbing the moisture in air, thus aforementioned developer becomes unstable etc.In addition, no matter wet system or responsibility, when preserving under the rayed conditions such as ultraviolet, aforementioned developer also can destabilization and produce nature colour developing, and this is also regarded as problem.
A large amount of methods has been had to be suggested and to adopt to solve the problem.
Such as, often adopt developer (and other reagent) freeze-drying in the test kit of liquid phase, and sealing is preserved.But such method requires complicated freeze-dry process step, and user in use, also need the operation steps adding solution redissolution.
Dry chemistry test bar has good stability, but its storage temperature is often room temperature, instead of the refrigerating temperature (2-8 DEG C) that test kit is conventional.Therefore, the developer stability problem in dry chemistry test bar is this area problem demanding prompt solution equally.Large quantifier elimination is also carried out to this both at home and abroad.
Such as a kind of method is: adopt tensio-active agent and the flavonoid system coloring matter with the alkyl of carbonatoms 8 ~ 16.But the method formula is complicated, and reagent is being difficult to buy on the market, and protective material itself has again certain interference to test result.
Also have by making methylenum coeruleum system developer and there is quaternary ammonium that carbonatoms is the alkyl of more than 12 or its salt coexists in liquid medium, make its stabilization.But the method to methylenum coeruleum system developer such as: two (dimethylamino) thiodiphenylamine of 10-(Carboxymethylaminocarbonyl)-3,7-.There is good stability but to common Trinder ' s developer and phenyl amines developer protectiveness poor or there is no stabilization.
Also research is had to adopt Ar-NH-NH-CO-NH
2stable reagent developer, or adopt a kind of color stability agent to have the reducing power stronger than developer, another kind of color stability agent has the reducing power between developer and the first color stability agent.But aforesaid method with the addition of stronger reducing substances, to the redox reaction of developer, there is certain interference effect, in order to by Interference Control in controlled range, higher to the requirement of protectant proportioning and concentration.Especially, when detecting analyte (creatinine, the transaminase) of low concentration, interference significantly.
Also have one of at least coexisting oxidation color former (developer) and Fructoamino-acid-oxidase (FAOD) and peroxidase (POD) in addition, make its stabilization.But the method can only stablize N-(carboxylic methylamino carbonyl)-4,4 '-two (dimethylin) pentanoic sodium not to be had stability action to other developers or acts on poor.
Aforesaid method all in the problem solving developer instability in varying degrees, but still also exists different defects, and therefore, further research is used for the stablizer of developer is very necessary.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is the stablizer and the application thereof that provide a kind of developer.Stablizer provided by the invention has satisfactory stability effect, even if still can play satisfactory stability effect to developer at a higher temperature, and does not disturb the color developing effect of developer.
The stablizer of developer provided by the invention, comprises azoic dyestuff;
Azoic dyestuff is selected from monoazo-dyes, disazo dyes or polyazo dye;
Monoazo-dyes is selected from No. l, Sudan red, No. 2, Sudan red, sudan orange B, it is red to lure, alizarin yellow R, orange G, lemon yellow, amaranth, Sunset yellow, Sudan red G, Eriochrome Red B, acid red 18 or crocein orange G;
Disazo dyes is selected from Xylene Red 66, Sudan black B, direct red 2B, alkaline brown g, Sudan red B, No. 3, Sudan red, No. 4, Sudan red or sudan red 7B;
Polyazo dye is direct black BN.
Azoic dyestuff is kind and the maximum dye of quantity in current Dyestuff Market, because gaining the name containing azo-group in its molecule.At present, azoic dyestuff is usually used in dyeing and the stamp of each fibrid, and painted for the dyeing such as leather, paper, soap, candle, timber, straw, feather and paint, ink, plastics, rubber, food etc.The present invention proves by experiment, and azoic dyestuff has stabilization to developer conventional in biochemistry detection, can improve the spontaneous colour developing of developer and make developer maintain better coloration ability.Further, azoic dyestuff as stablizer can also reduce to test effect interference.
In certain embodiments, azoic dyestuff is the composition of disazo dyes and polyazo dye, and wherein, the mol ratio of disazo dyes and polyazo dye is 15:40.
In certain embodiments, azoic dyestuff is the composition of monoazo-dyes and disazo dyes, and wherein, the mol ratio of monoazo-dyes and disazo dyes is 7:(4 ~ 20).
In an embodiment of the present invention, azoic dyestuff be selected from that crocein orange G, orange G, alkaline brown g, lemon yellow, alizarin yellow R, sudan orange B, Sudan black B, No. 3, Sudan red, No. 4, Sudan red, sudan red 7B, Sudan red B, Sudan red G, acid red 18, Xylene Red 66, Eriochrome Red B, temptation are red, direct black BN or direct red 2B
In an embodiment of the present invention, azoic dyestuff be selected from Sudan hong Ⅰ, No. 2, Sudan red, lure red, amaranth, Sudan red G, Eriochrome Red B, acid red 18, Xylene Red 66, direct red 2B, Sudan red B, No. 3, Sudan red, No. 4, Sudan red or sudan red 7B.
In an embodiment of the present invention, azoic dyestuff is selected from No. 3, Sudan red, No. 4, Sudan red, sudan red 7B, Sudan red B, Sudan red G, acid red 18, Xylene Red 66, Eriochrome Red B, direct red 2B or lures red.
In an embodiment of the present invention, the stablizer of developer also comprises any one in reductive agent, sequestrant, buffer reagent, auxiliary stabilizer, tensio-active agent or the combinations more than both.
The material that the valency that reducing substances refers to can reduce in oxygenant in chemical reaction the element playing oxygenizement reduces.Experiment shows, when stablizer provided by the invention and reducing substances with the use of time, the stability of developer can further improve.
In an embodiment of the present invention, the concentration of reductive agent is 10 μm of ol/L ~ 100 μm ol/L;
Reductive agent is selected from 1-Carbaphen, 1,5-diphenyl urea, N, N, N-trimethylamino urea, 4-Carbaphen, dihydroxyphenyl propane, gallic acid, carbohydrazide, paracetamol or gentisinic acid.
In certain embodiments, reductive agent is 1-Carbaphen, 4-Carbaphen, carbohydrazide or paracetamol.
In an embodiment of the present invention, the concentration of sequestrant is 20 μm of ol/L ~ 150 μm ol/L;
Sequestrant is selected from ethylenediamine tetraacetic acid (EDTA), diethylene triaminepentaacetic acid(DTPA), CDTA, O, O '-two (2-aminoethyl) ethylene glycol-N, N, N ', N '-tetraacethyl or nitrilotriacetic acid(NTA).
In certain embodiments, sequestrant is EDTA, GEDTA or DTPA.
In an embodiment of the present invention, the concentration of buffer reagent is 0.1mol/L;
Buffer reagent is selected from citric acid buffer agent, phosphate buffer, acetate buffer agent or Good ' s buffer reagent.
In certain embodiments, buffer reagent is Hepes damping fluid, phosphate buffer, MOPS damping fluid or MES damping fluid.
In an embodiment of the present invention, the massfraction of tensio-active agent is 0.5% ~ 1%;
Described tensio-active agent is selected from Tween series, Triton series, Brij is serial or Pluronic is serial.
In certain embodiments, tensio-active agent is TritonX100, PluronicF-68 or Tween20.
In an embodiment of the present invention, the massfraction of auxiliary stabilizer is 1% ~ 2%;
Auxiliary stabilizer is selected from carbohydrate or protein.
In certain embodiments, carbohydrate is sucrose, trehalose or sorbyl alcohol; Albumen is BSA.
In an embodiment of the present invention, developer is selected from phenyl amines, phenols, Trinder, connection nitrogen class or imidazoles.
In certain embodiments, phenyl amines developer is selected from TOPS (N-ethyl-N-(3-sulfopropyl)-3-monomethylaniline sodium salt), MAOS (N-ethyl-N-(2-hydroxyl-3-sulfopropyl)-3-5-dimethoxyaniline), MADB [N, two (4-sulphur the butyl)-3-5-xylidine of N-], TMB (3,3 ', 5,5 '-tetramethyl benzidine) or TOOS [N-ethyl-N-TOOS].
In certain embodiments, Trinder developer is selected from 4AA (4-AA), TBHBA (TBHBA) or DHBS (DBHS).
In certain embodiments, joining nitrogen class developer is ABTS [(2,2 '-azo-bis-(3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) di-ammonium salts)].
In certain embodiments, developer is selected from TMB, 4AA, MAOS, MADB, DHBS or TBHBA.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, tensio-active agent and buffer reagent, and wherein the concentration of azoic dyestuff is 1 μm of ol/L ~ 110 μm ol/L; The massfraction of tensio-active agent is 1%; The concentration of damping fluid is 0.1mol/L.
As preferably, azoic dyestuff is selected from sudan orange B, alizarin yellow R, crocein orange G, orange G, Sunset yellow or lemon yellow; Tensio-active agent is TritonX100; Damping fluid is MOPS damping fluid.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 1 μm of ol/L, and massfraction is the tensio-active agent of 1%, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the sudan orange B that concentration is 1 μm of ol/L, and massfraction is the TritonX100 of 1%, and concentration is the MOPS damping fluid of 0.1mol/L.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 110 μm of ol/L, and massfraction is the tensio-active agent of 1%, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the alizarin yellow R that concentration is 10 μm of ol/L, and concentration is the crocein orange G of 100 μm of ol/L, massfraction is the TritonX100 of 1%, and concentration is the MOPS damping fluid of 0.1mol/L.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 25 μm of ol/L, and massfraction is the tensio-active agent of 1%, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the orange G that concentration is 5 μm of ol/L, and concentration is the Sunset yellow of 20 μm of ol/L, massfraction is the TritonX100 of 1%, and concentration is the MOPS damping fluid of 0.1mol/L.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 5 μm of ol/L, and massfraction is the tensio-active agent of 1%, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the lemon yellow that concentration is 5 μm of ol/L, and massfraction is the TritonX100 of 1%, and concentration is the MOPS damping fluid of 0.1mol/L.
As preferably, stable for staining agent TMB of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TMB is 1:10000,110:10000,25:10000 or 5:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 7.4.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff and buffer reagent, and wherein the concentration of azoic dyestuff is 0.1 μm of ol/L ~ 1mmol/L; The concentration of damping fluid is 0.1mol/L.
As preferably, azoic dyestuff is selected from Xylene Red 66, Sudan black B, alkaline brown g, direct black BN, Sudan red B or sudan red 7B; Damping fluid is phosphate buffered saline buffer.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 0.1 μm of ol/L, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the Xylene Red 66 that concentration is 0.1 μm of ol/L, and concentration is the phosphate buffered saline buffer of 0.1mol/L.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 50 μm of ol/L, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the Sudan black B that concentration is 50 μm of ol/L, and concentration is the phosphate buffered saline buffer of 0.1mol/L.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 55 μm of ol/L, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the alkaline brown g that concentration is 15 μm of ol/L, and concentration is the direct black BN of 40 μm of ol/L, and concentration is the phosphate buffered saline buffer of 0.1mol/L.
As preferably, the stablizer of staining agent comprises the azoic dyestuff that concentration is 1075 μm of ol/L, and concentration is the damping fluid of 0.1mol/L.
Preferably, the stablizer of staining agent comprises the Sudan red B that concentration is 15 μm of ol/L, and concentration is the sudan red 7B of 1000 μm of ol/L, and concentration is the phosphate buffered saline buffer of 0.1mol/L.
As preferably, stable for staining agent 4AA and MAOS of the stablizer provided in this embodiment.
In this embodiment, the mole number of azoic dyestuff is 0.1:20000,50:20000,55:20000 or 1075:20000 with the ratio of the mole number sum of staining agent 4AA and MAOS.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.5.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, reductive agent, auxiliary stabilizer and buffer reagent, wherein the concentration of azoic dyestuff is 10 μm of ol/L ~ 15 μm ol/L, the concentration of reductive agent is 50 μm of ol/L ~ 100 μm ol/L, the massfraction of auxiliary stabilizer is 2%, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is selected from direct red 2B, it is red to lure; Reductive agent is 4-Carbaphen; Auxiliary stabilizer is sucrose; Damping fluid is HEPES damping fluid.
As preferably, the stablizer of staining agent comprises the direct red 2B that concentration is 10 μm of ol/L; Concentration is the 4-Carbaphen of 50 μm of ol/L; Massfraction is the sucrose of 2%; Concentration is the HEPES damping fluid of 0.1mol/L.
As preferably, azoic dyestuff be selected from for direct red 2B, lure red; Reductive agent is 4-Carbaphen; Auxiliary stabilizer is sucrose; Damping fluid is HEPES damping fluid.
As preferably, it is that the temptation of 15 μm of ol/L is red that the stablizer of staining agent comprises concentration; Concentration is the 4-Carbaphen of 100 μm of ol/L; Massfraction is the sucrose of 2%; Concentration is the HEPES damping fluid of 0.1mol/L.
As preferably, stable for staining agent 4AA and MADB of the stablizer provided in this embodiment.
In this embodiment, the mole number of azoic dyestuff is 10:20000 or 15:20000 with the ratio of the mole number sum of staining agent 4AA and MADB.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 7.5.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, sequestrant, auxiliary stabilizer and buffer reagent, wherein, the concentration of azoic dyestuff is 20 μm of ol/L, the concentration of sequestrant is 40 μm of ol/L, the massfraction of auxiliary stabilizer is 2%, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is No. 3, Sudan red; Sequestrant is EDTA; Auxiliary stabilizer is sucrose; Buffer reagent is HEPES.
Preferably, dying-stable agent comprises No. 3, the Sudan red that concentration is 20 μm of ol/L; Concentration is the EDTA of 40 μm of ol/L; Massfraction is the sucrose of 2%; Concentration is the HEPES of 0.1mol/L.
As preferably, stable for staining agent 4AA and MADB of the stablizer provided in this embodiment.
In this embodiment, the mole number of azoic dyestuff is 20:20000 with the ratio of the mole number sum of staining agent 4AA and MADB.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 7.5.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, sequestrant, auxiliary stabilizer and buffer reagent, wherein, the concentration of azoic dyestuff is 35 μm of ol/L, the massfraction of auxiliary stabilizer is 2%, the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is No. 4, Sudan red; Auxiliary stabilizer is sucrose; Buffer reagent is HEPES.
Preferably, dying-stable agent comprises No. 4, the Sudan red that concentration is 35 μm of ol/L; Massfraction is the sucrose of 2%; Concentration is the HEPES damping fluid of 0.1mol/L.
As preferably, stable for staining agent 4AA and MADB of the stablizer provided in this embodiment.
In this embodiment, the mole number of azoic dyestuff is 35:20000 with the ratio of the mole number sum of staining agent 4AA and MADB.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 7.5.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, reductive agent, sequestrant, tensio-active agent, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 55 μm of ol/L ~ 100 μm ol/L, the concentration of reductive agent is 10 μm of ol/L ~ 50 μm ol/L, the concentration of sequestrant is 100 μm of ol/L ~ 140 μm ol/L, the massfraction of tensio-active agent is 0.5%, the massfraction of auxiliary stabilizer is 1%, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is selected from direct red 2B, No. 3, Sudan red or Sudan red G; Reductive agent is carbohydrazide; Sequestrant is DTPA; Tensio-active agent is PluronicF-68; Auxiliary stabilizer is sorbyl alcohol; Damping fluid is MES damping fluid.
Preferably, the stablizer of developer comprises the direct red 2B that concentration is 100 μm of ol/L, and concentration is the carbohydrazide of 10 μm of ol/L, concentration is the DTPA of 100 μm of ol/L, massfraction is the PluronicF-68 of 0.5%, and massfraction is the sorbyl alcohol of 1%, and concentration is the MES damping fluid of 0.1mol/L.
Preferably, the stablizer of developer comprises No. 3, the Sudan red that concentration is 35 μm of ol/L, concentration is the Sudan red G of 20 μm of ol/L, concentration is the carbohydrazide of 50 μm of ol/L, concentration is the DTPA of 140 μm of ol/L, massfraction is the PluronicF-68 of 0.5%, and massfraction is the sorbyl alcohol of 1%, and concentration is the MES damping fluid of 0.1mol/L.
As preferably, stable for staining agent TMB of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TMB is 100:10000 or 55:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.0.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, reductive agent, tensio-active agent, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 45 μm of ol/L, and the concentration of reductive agent is 25 μm of ol/L, and the massfraction of tensio-active agent is 0.5%, and the massfraction of auxiliary stabilizer is 1%, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is acid red 18 and Eriochrome Red B, and reductive agent is carbohydrazide; Tensio-active agent is PluronicF-68; Auxiliary stabilizer is sorbyl alcohol; Damping fluid is MES damping fluid.
Preferably, the stablizer of developer comprises the acid red 18 that concentration is 40 μm of ol/L, concentration is the Eriochrome Red B of 5 μm of ol/L, concentration is the carbohydrazide of 25 μm of ol/L, massfraction is the PluronicF-68 of 0.5%, massfraction is the sorbyl alcohol of 1%, and concentration is the MES damping fluid of 0.1mol/L.
As preferably, stable for staining agent TMB of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TMB is 45:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.0.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, tensio-active agent, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 40 μm of ol/L, and the massfraction of tensio-active agent is 0.5%, and the massfraction of auxiliary stabilizer is 1%, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is acid red 18; Tensio-active agent is PluronicF-68; Auxiliary stabilizer is sorbyl alcohol; Damping fluid is MES damping fluid.
Preferably, the stablizer of developer comprises the acid red 18 that concentration is 40 μm of ol/L, and massfraction is the PluronicF-68 of 0.5%, and massfraction is the sorbyl alcohol of 1%, and concentration is the MES damping fluid of 0.1mol/L.
As preferably, stable for staining agent TMB of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TMB is 40:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.0.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, reductive agent, sequestrant and buffer reagent; Wherein, the concentration of azoic dyestuff is 35 μm of ol/L ~ 80 μm ol/L, and the concentration of reductive agent is 15 μm of ol/L ~ 30 μm ol/L, and the concentration of sequestrant is 80 μm of ol/L ~ 100 μm ol/L, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is selected from lemon yellow, orange G or Eriochrome Red B; Reductive agent is paracetamol; Sequestrant is GEDTA; Damping fluid is phosphate buffered saline buffer.
Preferably, the stablizer of developer comprises the lemon yellow that concentration is 60 μm of ol/L; Concentration is the orange G of 20 μm of ol/L; Concentration is the paracetamol of 30 μm of ol/L; Concentration is the GEDTA of 80 μm of ol/L; Concentration is the phosphate buffered saline buffer of 0.1mol/L.
Preferably, the stablizer of developer comprises the Eriochrome Red B that concentration is 15 μm of ol/L; Concentration is the orange G of 20 μm of ol/L; Concentration is the paracetamol of 15 μm of ol/L; Concentration is the GEDTA of 100 μm of ol/L; Concentration is the phosphate buffered saline buffer of 0.1mol/L.
As preferably, stable for staining agent TMB of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TMB is 80:10000 or 35:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.0.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, reductive agent and buffer reagent; Wherein, the concentration of azoic dyestuff is 75 μm of ol/L, and the concentration of reductive agent is 20 μm of ol/L, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is sudan red 7B and acid red 18; Reductive agent is paracetamol; Damping fluid is phosphate buffered saline buffer.
Preferably, the stablizer of developer comprises the sudan red 7B that concentration is 35 μm of ol/L; Concentration is the acid red 18 of 40 μm of ol/L; Concentration is the paracetamol of 20 μm of ol/L; Concentration is the phosphate buffered saline buffer of 0.1mol/L.
As preferably, stable for staining agent TMB of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TMB is 75:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.0.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, sequestrant and buffer reagent; Wherein, the concentration of azoic dyestuff is 40 μm of ol/L, and the concentration of sequestrant is 150 μm of ol/L, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is acid red 18; Sequestrant is GEDTA; Damping fluid is phosphate buffered saline buffer.
Preferably, the stablizer of developer comprises the acid red 18 that concentration is 40 μm of ol/L; Concentration is the GEDTA of 150 μm of ol/L; Concentration is the phosphate buffered saline buffer of 0.1mol/L.
As preferably, stable for staining agent TMB of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TMB is 40:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.0.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 55 μm of ol/L ~ 115 μm ol/L, and the massfraction of auxiliary stabilizer is 2%, and the concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is lemon yellow and orange G; Auxiliary stabilizer is BSA; Damping fluid is MES damping fluid.
Preferably, the stablizer of developer comprises the lemon yellow that concentration is 55 μm of ol/L; Massfraction is the BSA of 2%; Concentration is the MES damping fluid of 0.1mol/L.
Preferably, the stablizer of developer comprises the lemon yellow that concentration is 100 μm of ol/L; Concentration is the orange G of 15 μm of ol/L; Massfraction is the BSA of 2%; Concentration is the MES damping fluid of 0.1mol/L.
As preferably, stable for staining agent DHBS of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent DHBS is 55:10000 or 115:1000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.5.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, reductive agent, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 100 μm of ol/L; The concentration of reductive agent is 50 μm of ol/L; The massfraction of auxiliary stabilizer is 2%; The concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is direct red 2B; Auxiliary stabilizer is BSA; Damping fluid is MES damping fluid.
Preferably, the stablizer of developer comprises the direct red 2B that concentration is 100 μm of ol/L; Concentration is the 1-Carbaphen of 50 μm of ol/L; Massfraction is the BSA of 2%; Concentration is the MES damping fluid of 0.1mol/L.
As preferably, stable for staining agent DHBS of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent DHBS is 100:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.5.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, sequestrant, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 150 μm of ol/L; The concentration of sequestrant is 20 μm of ol/L; The massfraction of auxiliary stabilizer is 2%; The concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is No. 3, Sudan red; Sequestrant is EDTA; Auxiliary stabilizer is BSA; Damping fluid is MES damping fluid.
Preferably, the stablizer of developer comprises No. 3, the Sudan red that concentration is 150 μm of ol/L; Concentration is the EDTA of 20 μm of ol/L; Massfraction is the BSA of 2%; Concentration is the MES damping fluid of 0.1mol/L.
As preferably, stable for staining agent DHBS of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent DHBS is 150:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 6.5.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, reductive agent, tensio-active agent, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 10 μm of ol/L; The concentration of reductive agent is 50 μm of ol/L; The massfraction of tensio-active agent is 0.5%; The massfraction of auxiliary stabilizer is 2%; The concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is Xylene Red 66; Reductive agent is 4-Carbaphen; Tensio-active agent is Twenn20; Auxiliary stabilizer is trehalose; Damping fluid is MOPS damping fluid.
Preferably, the stablizer of developer comprises the Xylene Red 66 that concentration is 10 μm of ol/L; Concentration is the 4-Carbaphen of 50 μm of ol/L; Massfraction is the Twenn20 of 0.5%; Massfraction is the trehalose of 2%; Concentration is the MOPS damping fluid of 0.1mol/L.
As preferably, stable for staining agent TBHBA of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TBHBA is 10:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 7.4.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, tensio-active agent, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 125 μm of ol/L; The massfraction of tensio-active agent is 0.5%; The massfraction of auxiliary stabilizer is 2%; The concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is direct red 2B and lemon yellow; Tensio-active agent is Twenn20; Auxiliary stabilizer is trehalose; Damping fluid is MOPS damping fluid.
Preferably, the stablizer of developer comprises the direct red 2B that concentration is 25 μm of ol/L; Concentration is the lemon yellow of 100 μm of ol/L; Massfraction is the Twenn20 of 0.5%; Massfraction is the trehalose of 2%; Concentration is the MOPS damping fluid of 0.1mol/L.
As preferably, stable for staining agent TBHBA of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TBHBA is 125:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 7.4.
In one embodiment of the invention, the stablizer of staining agent comprises azoic dyestuff, tensio-active agent, auxiliary stabilizer and buffer reagent; Wherein, the concentration of azoic dyestuff is 85 μm of ol/L; The massfraction of tensio-active agent is 0.5%; The massfraction of auxiliary stabilizer is 2%; The concentration of buffer reagent is 0.1mol/L.
As preferably, azoic dyestuff is Sunset yellow; Tensio-active agent is Twenn20; Auxiliary stabilizer is trehalose; Damping fluid is MOPS damping fluid.
Preferably, the stablizer of developer comprises the Sunset yellow that concentration is 85 μm of ol/L; Massfraction is the Twenn20 of 0.5%; Massfraction is the trehalose of 2%; Concentration is the MOPS damping fluid of 0.1mol/L.
As preferably, stable for staining agent TBHBA of the stablizer provided in this embodiment.
In this embodiment, the mol ratio of azoic dyestuff and staining agent TBHBA is 85:10000.
In this embodiment, the suitable services pH value of the stablizer of staining agent is 7.4.
In an embodiment of the present invention, the mass ratio of the stablizer of developer and developer provided by the invention is 1:(10 ~ 100000).
In certain embodiments, the mass ratio of the stablizer of developer and developer provided by the invention is 1:(100 ~ 10000).
The application of stablizer in preparation biochemistry detection reagent of developer provided by the invention; Biochemistry detection is to the quantitative of bilirubin, oxyphorase, glucose, creatinine, uric acid, cholesterol, triglyceride, high density lipoprotein cholesterol, gpt or glutamic-oxal(o)acetic transaminase or qualitative detection.
Need to add corresponding oxydase, to produce hydrogen peroxide and above-mentioned chromogenic reagent when measuring above-mentioned analyte.Such as, glucose oxidase is added when measuring glucose.
Stablizer of the present invention also can be applied to dry chemistry and detect.Composition of the present invention is fixed on solid phase carrier, then dry.During test, sample is added on solid phase carrier, reacts.Satisfactory stability can be obtained equally.
The present invention adopts azoic dyestuff as the stablizer of developer, not only serves satisfactory stability effect, and can not cause interference to detection.Experiment shows, states adding of azoic dyestuff, improves the spontaneous colour developing of developer and makes developer maintain better coloration ability.Do not adding in the controlled trial of azoic dyestuff, staining agent all there occurs color change to a certain degree, and light absorption value also there occurs obvious rising, and after adopting stablizer provided by the invention, dye color changes not obvious, and light absorption value change is not remarkable.Further, after high temperature is placed for a long time, still good Detection results can be obtained.
Embodiment
The invention provides a kind of stablizer and application thereof of developer, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can change methods and applications herein or suitably changes and combination not departing from content of the present invention, spirit and scope, realizes and applies the technology of the present invention.
The reagent that the present invention adopts or instrument are all common commercially available product, all can buy in market.
Below in conjunction with embodiment, set forth the present invention further:
Embodiment 1
Be formulated as follows the composition shown in table:
Table 1 embodiment 1 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| TMB | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Sudan orange B | 1μmol/L | ||||
| Alizarin yellow R | 10μmol/L | ||||
| Orange G | 5μmol/L | ||||
| Lemon yellow | 5μmol/L | ||||
| Luo Yellow | 20μmol/L | ||||
| Crocein orange G | 100μmol/L | ||||
| TritonX100 | 1％ | 1％ | 1％ | 1％ | 1％ |
| MOPS | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
By hydrochloric acid or sodium hydroxide adjust ph to 7.4.
After each solution preparation, the absorbancy of 650nm is 0.02.Place 7 days for 50 DEG C, test the absorbancy of the 650nm of above-mentioned solution, result is as shown in the table:
The each scheme of table 2 embodiment 1 50 DEG C places absorbancy after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbancy | 0.201 | 0.156 | 0.143 | 0.135 | 0.137 |
In addition, get above-mentioned solution 1ml, add the HRP solution 100 μ l of 20U/L, add the hydrogen peroxide 100 μ l of 10mmol/L, 37 DEG C of hatching 3min, observe the color of above-mentioned solution, result is as shown in the table:
The each scheme of table 3 embodiment 1 50 DEG C places color after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | Blue | Blue | Blue | Blue |
Visible, adding of above-mentioned dyestuff, improve the spontaneous colour developing of TMB and make developer maintain better coloration ability.
Embodiment 2
Be formulated as follows the composition shown in table:
Table 4 embodiment 2 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| 4AA | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| MAOS | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Xylene Red 66 | -- | 0.1μmol/L | |||
| Sudan black B | -- | 50μmol/L | |||
| Alkaline brown g | -- | 15μmol/L | |||
| Sudan red B | -- | 75μmol/L | |||
| Direct black BN | -- | 40μmol/L | |||
| Sudan red 7B | -- | 1mmol/L | |||
| Phosphoric acid buffer | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
By hydrochloric acid or sodium hydroxide adjust ph to 6.5.
After each solution preparation, the absorbancy of 630nm is 0.015.Place 7 days for 60 DEG C, test the absorbancy of the 630nm of above-mentioned solution, result is as shown in the table:
The each scheme of table 5 embodiment 2 60 DEG C places absorbancy after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbancy | 0.101 | 0.045 | 0.063 | 0.071 | 0.083 |
In addition, get above-mentioned solution 1ml, add the HRP solution 100 μ l of 20U/L, add the hydrogen peroxide 100 μ l of 10mmol/L, 37 DEG C of hatching 3min, observe the color of above-mentioned solution, result is as shown in the table:
The each scheme of table 6 embodiment 2 60 DEG C places color after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | Blue | Blue | Blue | Blue |
Visible, adding of above-mentioned dyestuff, improve the spontaneous colour developing of 4AA and MAOS and make developer maintain better coloration ability.
Embodiment 3
Be formulated as follows the composition shown in table:
Table 7 embodiment 3 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| 4AA | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| MADB | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Direct red 2B | -- | 10μmol/L | |||
| No. 3, Sudan red | -- | 20μmol/L | |||
| No. 4, Sudan red | -- | 35μmol/L | |||
| Lure red | -- | 15μmol/L | |||
| EDTA | -- | 40μmol/L | |||
| 4-Carbaphen | -- | 50μmol/L | 100μmol/L | ||
| Sucrose | 2％ | 2％ | 2％ | 2％ | 2％ |
| HEPES | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
By hydrochloric acid or sodium hydroxide adjust ph to 7.5.
After each solution preparation, the absorbancy of 630nm is 0.014.Place 7 days for 60 DEG C, test the absorbancy of the 630nm of above-mentioned solution, result is as shown in the table:
The each scheme of table 8 embodiment 3 60 DEG C places absorbancy after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbancy | 0.125 | 0.035 | 0.045 | 0.077 | 0.047 |
In addition, get above-mentioned solution 1ml, add the HRP solution 100 μ l of 20U/L, add the hydrogen peroxide 100 μ l of 10mmol/L, 37 DEG C of hatching 3min, observe the color of above-mentioned solution, result is as shown in the table:
The each scheme of table 9 embodiment 3 60 DEG C places color after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | Blue | Blue | Blue | Blue |
Visible, adding of above-mentioned dyestuff, improve the spontaneous colour developing of 4AA and MADB and make developer maintain better coloration ability.
Embodiment 4
Be formulated as follows the composition shown in table:
Table 10 embodiment 4 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| TMB | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Direct red 2B | 100μmol/L | ||||
| No. 3, Sudan red | 35μmol/L | ||||
| Acid red 18 | 40μmol/L | 40μmol/L | |||
| Eriochrome Red B | 5μmol/L | ||||
| Sudan red G | 20μmol/L | ||||
| DTPA | 100μmol/L | 150μmol/L | 100μmol/L | ||
| Carbohydrazide | 10μmol/L | 25μmol/L | 50μmol/L | ||
| Sorbyl alcohol | 1％ | 1％ | 1％ | 1％ | 1％ |
| Pluronic F-68 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 |
| MES | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
By hydrochloric acid or sodium hydroxide adjust ph to 6.0.
After each solution preparation, the absorbancy of 650nm is 0.018.Place 7 days for 70 DEG C, test the absorbancy of the 650nm of above-mentioned solution, result is as shown in the table:
The each scheme of table 11 embodiment 4 70 DEG C places absorbancy after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbancy | 0.205 | 0.103 | 0.095 | 0.085 | 0.071 |
In addition, get above-mentioned solution 1ml, add the HRP solution 100 μ l of 20U/L, add the hydrogen peroxide 100 μ l of 10mmol/L, 37 DEG C of hatching 3min, observe the color of above-mentioned solution, result is as shown in the table:
The each scheme of table 12 embodiment 4 70 DEG C places color after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | Blue | Blue | Blue | Blue |
Visible, adding of above-mentioned dyestuff, improve the spontaneous colour developing of TMB and make developer maintain better coloration ability.
Embodiment 5
Be formulated as follows the composition shown in table:
Table 13 embodiment 5 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| TMB | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Lemon yellow | 60μmol/L | ||||
| Sudan red 7B | 35μmol/L | ||||
| Acid red 18 | 40μmol/L | 40μmol/L | |||
| Eriochrome Red B | 15μmol/L | ||||
| Orange G | 20μmol/L | 20μmol/L | |||
| GEDTA | 80μmol/L | 100μmol/L | 150μmol/L | ||
| Paracetamol | 30μmol/L | 20μmol/L | 15μmol/L | ||
| Phosphate buffer | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
By hydrochloric acid or sodium hydroxide adjust ph to 6.0.
After each solution preparation, the absorbancy of 650nm is 0.019.Place 7 days for 60 DEG C, test the absorbancy of the 650nm of above-mentioned solution, result is as shown in the table:
The each scheme of table 14 embodiment 5 60 DEG C places absorbancy after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Absorbancy | 0.205 | 0.030 | 0.045 | 0.025 | 0.021 |
In addition, get above-mentioned solution 1ml, add the HRP solution 100 μ l of 20U/L, add the hydrogen peroxide 100 μ l of 10mmol/L, 37 DEG C of hatching 3min, observe the color of above-mentioned solution, result is as shown in the table:
The each scheme of table 15 embodiment 5 60 DEG C places color after 7 days
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| Color | Pale blue | Blue | Blue | Blue | Blue |
Visible, adding of above-mentioned dyestuff, improve the spontaneous colour developing of TMB and make developer maintain better coloration ability.
Embodiment 6
Be formulated as follows the composition shown in table:
Table 16 embodiment 6 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| DHBS | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Sterol esterase | 200KU/L | 200KU/L | 200KU/L L | 200KU/L | 200KU/L |
| RCO | 300KU/L | 300KU/L | 300KU/L | 300KU/L | 300KU/L |
| Peroxidase | 300KU/L | 300KU/L | 300KU/L | 300KU/L | 300KU/L |
| Direct red 2B | -- | 100μmol/L | |||
| No. 3, Sudan red | -- | 150μmol/L | |||
| Lemon yellow | -- | 55μmol/L | 100μmol/L | ||
| Orange G | -- | 15μmol/L | |||
| EDTA | -- | 20μmol/L | |||
| 1-Carbaphen | -- | 50μmol/L | |||
| BSA | 2％ | 2％ | 2％ | 2％ | 2％ |
| MES | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
By hydrochloric acid or sodium hydroxide adjust ph to 6.5.
Above-mentioned 5 schemes respectively place portion respectively in 2-8 DEG C and 45 DEG C of temperature, respectively with solution testing 2.5mmol/L, 8.7mmol/L, 11mmol/L total cholesterol serum sample that in 5 schemes, 2-8 DEG C is preserved (above-mentioned total cholesterol concentration adopts full automatic biochemical apparatus to measure) after 3 days, obtain absorbancy.Obtain regression equation according to concentration of specimens drawing standard curve.
Testing method is add 0.3ml serum sample at the above-mentioned solution of 2.7ml, and 37 DEG C of hatching 5min, read absorbancy at 520nm.
Above-mentioned 5 schemes are tested above-mentioned 3 samples after the same method at 60 DEG C of storage solutions, brings absorbancy result into above-mentioned regression equation, calculate test sample concentration.Result (unit: mmol/L) as shown in the table:
The each scheme absorbancy of table 17 embodiment 6
| Sample actual concentrations | Option A | Option b | Scheme C | Scheme D | Scheme E |
| 2.5mmol/L | 3.5 | 2.8 | 2.7 | 2.9 | 2.7 |
| 8.7mmol/L | 6.3 | 8.2 | 8.2 | 7.8 | 7.9 |
| 11mmol/L | 7.5 | 10.5 | 3.7 | 10.1 | 10.2 |
As can be seen from the above table, do not add the protectant option A of the present invention, low value (2.5mmol/L) is higher, is to cause because background raises.And high level (11mmol/L) reduction loses coloration ability because developer is rotten.And after adding protective material of the present invention, the problem of these two aspects is obtained for significant improvement, namely improve the stability of developer.
Embodiment 7
Be formulated as follows the composition shown in table:
Table 18 embodiment 7 experimental program
| Option A | Option b | Scheme C | Scheme D | Scheme E | |
| TBHBA | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L | 10mmol/L |
| Urico-oxidase | 200KU/L | 200KU/L | 200KU/L L | 200KU/L | 200KU/L |
| Peroxidase | 300KU/L | 300KU/L | 300KU/L | 300KU/L | 300KU/L |
| Xylene Red 66 | -- | 10μmol/L | |||
| Direct red 2B | -- | 25μmol/L | |||
| Lemon yellow | -- | 100μmol/L | L | ||
| Luo Yellow | -- | 85μmol/L | |||
| EDTA | -- | 20μmol/L | |||
| 4-Carbaphen | -- | 50μmol/L | |||
| Trehalose | 2％ | 2％ | 2％ | 2％ | 2％ |
| Twenn 20 | 0.5％ | 0.5％ | 0.5％ | 0.5％ | 0.5％ |
| MOPS | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L | 0.1mol/L |
By hydrochloric acid or sodium hydroxide adjust ph to 7.4.
Be coated on the filter paper of 1cm × 1cm by above-mentioned 5 schemes, each scheme is coated with 2 filter paper.At 40 DEG C of baking 2h.After oven dry, two filter paper are put into respectively the temperature of 2-8 DEG C and 55 DEG C, after 7 days.Respectively with filter paper test 0.11mmol/L, 0.67mmol/L, 1.1mmol/L uric acid serum sample (above-mentioned uric acid concentration adopts full automatic biochemical apparatus to measure) that in 5 schemes, 2-8 DEG C is preserved, obtain reflectivity.Obtain regression equation according to concentration of specimens drawing standard curve.
Testing method for drip 30 μ l serum samples on filter paper, and 37 DEG C of hatching 3min, read 530nm and read reflectivity.
Above-mentioned 5 schemes are stored filter paper at 60 DEG C and tests above-mentioned 3 samples after the same method, bring reflectivity results into above-mentioned regression equation, calculate test sample concentration.Result (unit: mmol/L) as shown in the table:
The each scheme of table 19 embodiment 7 measures concentration
| Sample actual concentrations | Option A | Option b | Scheme C | Scheme D | Scheme E |
| 0.11mmol/L | 0.20 | 0.13 | 0.14 | 0.13 | 0.12 |
| 0.67mmol/L | 0.55 | 0.65 | 0.66 | 0.62 | 0.68 |
| 1.1mmol/L | 0.75 | 1.02 | 1.05 | 1.03 | 1.12 |
As can be seen from the above table, do not add the protectant option A of the present invention, low value (0.11mmol/L) is higher, is to cause because background raises.And high level (1.1mmol/L) reduction loses coloration ability because developer is rotten.And after adding protective material of the present invention, the problem of these two aspects is obtained for significant improvement, namely improve the stability of developer.
Below be only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a stablizer for developer, is characterized in that, comprises azoic dyestuff;
Described azoic dyestuff is selected from monoazo-dyes, disazo dyes or polyazo dye;
Described monoazo-dyes is selected from No. l, Sudan red, No. 2, Sudan red, sudan orange B, it is red to lure, alizarin yellow R, orange G, lemon yellow, amaranth, Sunset yellow, Sudan red G, Eriochrome Red B, acid red 18 or crocein orange G;
Described disazo dyes is selected from Xylene Red 66, Sudan black B, direct red 2B, alkaline brown g, Sudan red B, No. 3, Sudan red, No. 4, Sudan red or sudan red 7B;
Described polyazo dye is direct black BN.
2. the stablizer of developer according to claim 1, is characterized in that, also comprises any one in reductive agent, sequestrant, buffer reagent, auxiliary stabilizer, tensio-active agent or both above combinations.
3. the stablizer of developer according to claim 2, is characterized in that, the concentration of wherein said reductive agent is 10 μm of ol/L ~ 100 μm ol/L;
Described reductive agent is selected from 1-Carbaphen, 1,5-diphenyl urea, N, N, N-trimethylamino urea, 4-Carbaphen, dihydroxyphenyl propane, gallic acid, carbohydrazide, paracetamol or gentisinic acid.
4. the stablizer of developer according to claim 2, is characterized in that, the concentration of wherein said sequestrant is 20 μm of ol/L ~ 150 μm ol/L;
Described sequestrant is selected from ethylenediamine tetraacetic acid (EDTA), diethylene triaminepentaacetic acid(DTPA), CDTA, O, O '-two (2-aminoethyl) ethylene glycol-N, N, N ', N '-tetraacethyl or nitrilotriacetic acid(NTA).
5. the stablizer of developer according to claim 2, is characterized in that, the concentration of wherein said buffer reagent is 0.1mol/L;
Described buffer reagent is selected from citric acid buffer agent, phosphate buffer, acetate buffer agent or Good ' s buffer reagent.
6. the stablizer of developer according to claim 2, is characterized in that, the massfraction of wherein said tensio-active agent is 0.5% ~ 1%;
Described tensio-active agent is selected from Tween series, Triton series, Brij is serial or Pluronic is serial.
7. the stablizer of developer according to claim 2, is characterized in that, the massfraction of wherein said auxiliary stabilizer is 1% ~ 2%;
Described auxiliary stabilizer is selected from carbohydrate or protein.
8. the stablizer of the developer according to any one of claim 1 ~ 7, is characterized in that, described developer is selected from phenyl amines, phenols, Trinder, connection nitrogen class or imidazoles.
9. the stablizer of the developer according to any one of claim 1 ~ 7, is characterized in that, the mass ratio of the stablizer of described developer and the developer described in any one of claim 1 ~ 7 is 1:(10 ~ 100000).
10. the application of stablizer in preparation biochemistry detection reagent of the developer described in any one of claim 1 ~ 7; Described biochemistry detection is to the quantitative of glucose, creatinine, uric acid, cholesterol, triglyceride, high density lipoprotein cholesterol, gpt or glutamic-oxal(o)acetic transaminase or qualitative detection.
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| CN106771130A (en) * | 2016-11-17 | 2017-05-31 | 陕西师范大学 | A kind of enzyme-linked immune analytic method |
| CN108918415A (en) * | 2018-07-19 | 2018-11-30 | 美康生物科技股份有限公司 | People's total antioxidant status detection kit |
| CN110791122A (en) * | 2019-11-28 | 2020-02-14 | 迈克生物股份有限公司 | Sudan black B staining kit |
| CN112029817A (en) * | 2020-07-13 | 2020-12-04 | 兰州百源基因技术有限公司 | Creatinine detection kit and use method thereof |
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